微生物转化浮萍资源生产蛋白酶的条件
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摘要
利用微生物转化富含蛋白质的浮萍生产蛋白酶,为浮萍的高值化利用开辟一条新途径。在摇瓶转化的基础上,探索在5L发酵罐上沙雷氏菌(Serratiasp.)SYBC H转化浮萍生产蛋白酶的最佳转化条件。通过对温度、通风量、搅拌转速、pH等参数的研究,发现在5L发酵罐上,沙雷氏菌SYBC H转化浮萍生产蛋白酶的最佳条件为通风量6vvm,搅拌转速400r/min,发酵温度30℃。转化过程中,采用全自动控制pH值(9.0),蛋白酶的产量比不控制pH的对照组提高了23.3%。进一步研究发现,利用微生物沙雷氏菌SYBC H转化浮萍生产的蛋白酶具有耐有机溶剂的特性,在某些亲水性有机溶剂中保留90%以上的活性,是一种稀少珍贵的极端酶。
Novel microbial transformation methods are explored for high-value utilization of protein-rich duckweed to produce proteases.Based on flask transformation,the transformation conditions for the production of proteases from duckweed by Serratiasp.SYBC H were optimized in a 5 Lfermentor.Through study on temperature,ventilation volume,stirring speed and pH,it is found that the best transformation conditions were ventilation volume 6 vvm,stirring speed 400 r/min,culture temperature 30 ℃.The protease production with constant auto-feed control of pH9.0 during process was 23.3% higher than that from pH non-control process.Further studies have shown that the protease produced from duckweed by Serratiasp.SYBC H still retained more than 90% activity in some hydrophilic orga-nic solvents,making it a rare and precious extreme protease.
Novel microbial transformation methods are explored for high-value utilization of protein-rich duckweed to produce proteases.Based on flask transformation,the transformation conditions for the production of proteases from duckweed by Serratiasp.SYBC H were optimized in a 5 Lfermentor.Through study on temperature,ventilation volume,stirring speed and pH,it is found that the best transformation conditions were ventilation volume 6 vvm,stirring speed 400 r/min,culture temperature 30 ℃.The protease production with constant auto-feed control of pH9.0 during process was 23.3% higher than that from pH non-control process.Further studies have shown that the protease produced from duckweed by Serratiasp.SYBC H still retained more than 90% activity in some hydrophilic orga-nic solvents,making it a rare and precious extreme protease.
引文
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[23]Ying J W,Zhang J,Zhang L N.Hazards and control of Cyanobacteria[J].Guangdong Chemical Industry,2017,44(11):212-213.应佳薇,张菁,张丽娜.关于蓝藻危害与防治[J].广东化工,2017,44(11):212-213.
[24]Zeng Z Q,Li X L,Shi Y J.Experimental study on the bacteriostasis of extract from blooma blue-green algae[J].Anti-Infection Pharmacy,2007,4(4):151-153.曾昭琪,李祥麟,石玉洁.水华蓝藻萃取物抑菌作用的实验研究[J].抗感染药学,2007,4(4):151-153.
[25]Li Y,Zhang J W,Wei J,et al.Advances in mechanism of the occurrence,hazard,and prevention/control utilization of cyanophytic blooms in China[J].Journal of Microbiology,2015,35(4):93-97.李媛,张家卫,魏杰,等.我国蓝藻水华的发生机理、危害及防控利用研究进展[J].微生物学杂志,2015,35(4):93-97.
[2]Kutschera U,Niklas K J,Darwin-wallace D.Survival of the fastest in populations of duckweeds and the evolutionary history of an enigmatic group of angiosperms[J].Plant Biol,2015,1(s1):24-32.
[3]Ziegler P,Adelmann K,Zimmer S,et al.Relative in vitro growth rates of duckweeds(Lemnaceae)—the most rapidly growing higher plants[J].Plant Biol,2015,17(s1):33.
[4]Falabi J A,Gerba C P,Karpiscak M M.Giardia and cryptosporidium removal from waste-water by a duckweed(Lemna gibba L.)covered pond[J].Lett Applied Microbiol,2002,34(5):384-387.
[5]Zhao Y,Fang Y,Jin Y.Pilot-scale comparison of four duckweed strains from different genera for potential application in nutrient recovery from waste water and valuable biomass[J].Plant Biol,2015,17(s1):82-90.
[6]Kabir A N M A,Hossain M A,Rahman M S.Use of duckweed as feed for fishes in polyculture[J].J Agr Rural Dev,2009,7(7):157-160.
[7]Landesman L,Chang J,Yamamoto Y,et al.Nutritional value of waste water-grown duckweed for fish and shrimp feed[J].World Aquacult,2002,33(4):39-40.
[8]Olshanskaya L N,Sobgaida N A,Tarushkina Y A.Effect of a magnetic field on extraction of heavy metals from waste water with duckweed[J].Chem Petrol Eng,2008,44(7):475-479.
[9]Sree K S,Muellerroeber B,Brandt R,et al.Phytotoxicity of cobalt ions on the duckweed Lemna minor—Morphology,ion uptake,and starch accumulation[J].Chemosphere,2015(131):149-156.
[10]Cui W,Cheng J J.Growing duckweed for biofuel production:a review[J].Plant Biol,2015,17(s1):16-23.
[11]Cheng J J,Stomp A M.Growing duckweed to recover nutrients from wastewaters and for production of fuel ethanol and animal feed[J].Clean-Soil Air Water,2009,37(1):17-26.
[12]Liu G,Wright M M,Zhao Q,et al.Catalytic fast pyrolysis of duckweed:effects of pyrolysis parameters and optimization of aromatic production[J].J Anal Appl Pyrol,2015(112):29-36.
[13]Gomes M P,Juneau P.Oxidative stress in duckweed(Lemna minor L)induced by glyphosate:is the mitochondrial electron transport chain a target of this herbicide?[J].Environ Pollut,2016(218):402-409.
[14]Casalta E,Aguera E,Picou C,et al.A comparison of laboratory and pilot-scale fermentation in wine making conditions[J].Appl Microbiol Biot,2010,87(5):1665-1673.
[15]Rahman R N,Mahamad S,Salleh A B,et al.A new organic solvent tolerant protease from Bacillus pumilus 115b[J].J Ind Microbiol Biotechnol,2007(34):509-517.
[16]Gupta A,Khara S K.A protease stable in organic solvents from solvent tolerant strain of Pseudomonas aeruginosa[J].Bioresour Technol,2006,97(15):1788-1793.
[17]Bhosale S H,Rao M B,Deshpande V,et al.Thermostability of high activity alkaline protease from Conidiobolus coronatus(NCI86.8.20)[J].Enzyme Microb Technol,1995,17(2):136-139.
[18]Tao J F,Lai Y T,Ren Y,et al.Preliminary study on fermentative process of Vibrio strain[J].Industrial Microorganism,2011,41(3):86-89陶家发,赖迎迢,任燕,等.弧菌菌株发酵工艺初步研究[J].工业微生物,2011,41(3):86-89.
[19]Li H,Xu H,Li S,et al.Effects of dissolved oxygen and shear stress on the synthesis and molecular weight of welan gum produced from Alcaligenes sp.CGMCC2428[J].Process Biochem,2011,46(11):1172-1178.
[20]Zhu Z L,Wang D,Xu Y.Improvement of fermentor facility to enhance liquid fermentation level of membrance-bound lipase produced by Rhizopus chinensis[J].Industrial Microorganism,2013,43(2):6-12.朱增亮,王栋,徐岩.发酵罐装置改进提高华根霉膜结合脂肪酶液态发酵水平[J].工业微生物,2013,43(2):6-12.
[21]Li G Y,Liao X R,Cai Y J,et al.Protease production by Serratiasp.SYBC H with blue algae[J].Chinese Journal of Environmental Engineering,2012,6(3):1043-1046.李桂英,廖祥儒,蔡宇杰,等.沙雷氏菌发酵蓝藻生产蛋白酶[J],环境工程学报,2012,6(3):1043-1046.
[22]Wang Y,Li Y,Feng Y,et al.Research progress on cyanobacterial toxins and the cyanotoxin synthetase gene[J].Journal of Ecology,2017,36(2):517-523.汪洋,李樾,冯悦,等.蓝藻毒素的类型及其产毒基因[J].生态学杂志,2017,36(2):517-523.
[23]Ying J W,Zhang J,Zhang L N.Hazards and control of Cyanobacteria[J].Guangdong Chemical Industry,2017,44(11):212-213.应佳薇,张菁,张丽娜.关于蓝藻危害与防治[J].广东化工,2017,44(11):212-213.
[24]Zeng Z Q,Li X L,Shi Y J.Experimental study on the bacteriostasis of extract from blooma blue-green algae[J].Anti-Infection Pharmacy,2007,4(4):151-153.曾昭琪,李祥麟,石玉洁.水华蓝藻萃取物抑菌作用的实验研究[J].抗感染药学,2007,4(4):151-153.
[25]Li Y,Zhang J W,Wei J,et al.Advances in mechanism of the occurrence,hazard,and prevention/control utilization of cyanophytic blooms in China[J].Journal of Microbiology,2015,35(4):93-97.李媛,张家卫,魏杰,等.我国蓝藻水华的发生机理、危害及防控利用研究进展[J].微生物学杂志,2015,35(4):93-97.